FAQ: Can longer or shorter incubation times be used?

Yes. For assembling 2-3 fragments, 15 minute incubation times are sufficient. For assembling 4-6 fragments, 60 minute incubation times are recommended. Reaction times less than 15 minutes are generally not recommended. Extended incubation times (up to 4 hours) have been shown to improve assembly efficiencies in some cases. Do not incubate the reaction overnight.